Int. J. Chem. Sci.: 10(2), 2012, 1028-1038 ISSN 0972-768X www.sadgurupublications.com

GC-MS ANALYSIS OF LEAF EXTRACTS OF PTEROSPERMUM CANESCENS Roxb. () K. P. JAIGANESH* and G. ARUNACHALAMa

Centre for Research and Development, PRIST University, THANJAVUR – 613403 (T.N.) INDIA aP.G. P. College of Pharmaceutical Sciences and Research Institute, NAMAKKAL – 637207 (T.N.) INDIA

ABSTRACT

Pterospermum canescens Roxb., is one of the medicinally important belonging to the family Sterculiaceae, is commonly known as “Sembolavu” in Tamil. Traditionally the plant is used for the treatment of headache, bronchitis, throat infections, skin diseases, leprosy, inflammation and sores. The present study was planned to identity the number of constituents present in Pterospermum canescens Roxb., leaf by quantitative GC-Ms analysis. The analysis was carried out for the petroleum ether, chloroform and methanol leaf extracts of Pterospermum canescens Roxb., and the results were exhibited the presence of 15, 6 and 20 compounds in the above respective extracts. The major chemical constituents are squalene, phytol, α-sitosterol, vitamin-E and tocopherol.

Key words: Pterosperumum canescens, Sterculiaceae, GC-MS, Phytol, Squalene.

INTRODUCTION

For thousands of years, plant products and their modified derivatives have been rich source for clinically useful drugs. Even today, about 80% of the world’s population relies predominantly on and plant extracts for health care1. Natural products as a pure compound or as a standardized plant extracts provide unlimited opportunities for new drug lead because of unmatched availability of chemical diversity. Therefore, there is a need to develop an efficient, safe and inexpensive drugs from plant source are of great importance2.

The Pterospermum Schreb. (Sterculiaceae) represents of about 40 species in the world, of which 12 species were reported from India and 8 species from Tamil Nadu state3. An ethno medicinal plant species Pterospermum canescens Roxb. (Syn:

______*Author for correspondence; E-mail: [email protected]; Mo.: +09942380275 Int. J. Chem. Sci.: 10(2), 2012 1029

Pterospermum suberifolium Lam.) is locally known as “Sembolavu” was distributed in all districts of Tamil Nadu. Ethno medicinally, the leaves are used for headache4 and Valiyan tribals from Dindigal district, applied leaf paste mixed with the stem bark of Drypetes roxburghii (Wall.) Hurus (Putranjivaceae, “Pilla maram”), a few leaves of Blepharis maderaspatensis (L.) B.Heyne (Acanthaceae, “Elumbu otti”) and the yellow yolks of 2 eggs on fractured bones5. Flowers and bark are charred, mixed with kamala and applied in suppurating small pox6. A pharmacological activity such as anti cancer activity, cytotoxicity has also been reported7,8. Hence, the objective of the present study is to identify the phytochemical constituents of Pterospermum canescans Roxb., with the aid of GC-MS technique.

EXPERIMENTAL Materials and methods Plant material

The leaves of Pterospermum canescens Roxb., were collected in the month of May from the Kalapet vicinity of Pondicherry and the collected plant material was botanically identified and confirmed by the plant Taxonomist Mr. A. C. Tangavelou and Dr. Susai, St. Joseph’s College (Autunomous), Tiruchirappalli, Tamil Nadu. A voucher specimen (KPJ 42) was prepared and deposited at the department for future reference. Preparation of extracts

The collected leaves were subjected to shade drying and pulverized to coarse powder in a mechanical grinder. The coarse leaf powder was successively extracted with various solvents such as petroleum ether (40o – 60oC), chloroform and methanol by using continuous hot percolation method with the help of Soxhlet apparatus9,10. The extracts were collected and distilled off on a water bath at atmospheric pressure and the last trace of the solvent was removed in vacuo. The resulted extracts were used for GC-MS analysis. GC-MS analysis of plant extracts11

The GC-MS analysis was carried out on a GC Clarus 500 Perkin Elmer system, comprising an Aoc-20i Auto sampler and Gas chromatograph interfaced to a mass spectrometer instrument, employing the conditions: Column Elite -5Ms fused silica capillary column (30 x 0.25 mm, 1D x 1mm df, composed of 95% Dimethyl Poly diloxane, 5% Diphenyl Polysixone), operating in electron impact mode at 70 ev; helium (99.999%) was used as carrier gas at a constant flow of 1 mL/min and an injection volume of 2 mL was employed (split ratio of 10 : 1), injector temperature is 200oC; ion-source temperature is 1030 K. P. Jaiganesh and G. Arunachalam: GC-MS Analysis of….

200oC. The oven temperature was programmed from 110oC (isothermal for 2 min), with an increase of 10oC /min, to 200oC, then 5oC /min to 280oC ending with a 9 min isothermal at 280oC. Mass spectra were taken at 70 ev; a scan interval of 0.5 seconds and fragments from 40 to 450 Da. Total GC running time is 36 min. Identification of compounds

GC-MS chromatogram of petroleum ether, chloroform and methanol leaf extracts of Pterospermum canescens Roxb., (Fig. 1-3), showed 15, 6 and 20 peaks indicating the presence of respective number of compounds. Interpretation of mass spectrum was conducted by using the database of National Institute of Standard and Technology (NIST) having more than 62,000 patterns. The name, molecular weight and structure of the components of test materials were characterized and identified (Table 1-3). Mass spectrum of some important compounds present in this plant was depicted (Fig. 4-13).

RESULTS AND DISCUSSION

GC-MS Chromatogram of petroleum ether leaf extract of Pterospermum canescens Roxb., (Fig. 1) showed 15 peaks represent the phyto constituents were α-sito sterol (19.38%), 3, 7, 11, 15-tetramethyl-2-hexa decane-1-ol (18-56%), ricinoleic acid (10.56%), vitamin-E (10.49%), phytol (9.71%), α-tocopherol (6.29%), diethyl phthalate (5.09%), squalene (5.07%), benzhydrazide-3-mthoxy-N2-(4-phenylcyclo hexylideno (4.62%), benzoic acid,4- heptyl-4-cyanophenyl ester (4.41%) and n-hexa decanoic acid (4.08%).

Fig. 1: Chromatogram of petroleum ether leaf extract

GC-Ms chromatogram of chloroform leaf extract of Pterospermum canescens Roxb., (Fig. 2) showed 6 peaks, represent the phyto constituents, were (3,7,7-trimethyl-1-penta-1,3- Int. J. Chem. Sci.: 10(2), 2012 1031 dienyl-2-oxabicyclo [3.2.0] hept-3-ene (49.04%), 2-allyl-4-methylphenol (30.49%) and asarone (9.43%).

Fig. 2: Chromatogram of chloroform leaf extract

GC-MS chromatogram of methanol leaf extract of Pterospermum canescens Roxb., (Fig. 3) shows 20 peaks, represent the phytoconstituents were diethyl phthalate (53.84%), ricinoleic acid (20.74%), glyceryl monoricinoleate (10.25%), methyl ricinoleate (3.93%), 4- ((1E)-3-hydroxy-1-propenyl)-2-methoxyphenol (2.33%) and 7-methyl-Z-tetradecen-1-ol acetate (2.10%).

Fig. 3: Chromatogram of methanol leaf extract

It is worthwhile, to indicate that there are no data in the literature of investigated the phytochemical constituents of Pterospermum canescens Roxb., leaves for comparison. Due to the limitation of present study, we could not evaluate the biological properties of Pterospermum canescens Roxb., are suggested for future research. 1032 K. P. Jaiganesh and G. Arunachalam: GC-MS Analysis of….

Table 1: Components identified in the petroleum ether leaf extract of Pterospermum canescens Roxb.

Molecular Peak S. No. RT Name of the compound MW formula area%

1 8.67 2(4H)-Benzofuranone,5,6,7,7α- C11H16O2 180 0.21 tetrahydro-4,4,7α-trimethyl-

2 9.23 Diethyl Phthalate C12H14O4 222 5.09

3 9.41 Dihydrojasmone C11H18O 166 0.42

4 11.38 3,3-Dimethyl-hepta-4,5-dien-2-ol C9H16O 140 0.28

5 11.93 3,7,11,15-Tetramethyl-2-hexadecen-1-ol C20H40O 296 18.56

6 13.50 n-Hexadecanoic acid C16H32O2 256 4.08

7 15.36 Phytol C20H40O 296 9.71

8 16.13 9-Octadecenal C18H34O 266 0.84

9 23.58 Benzoicacid,4-heptyl-4-cyanophenyl ester C21H23NO2 321 4.41

10 23.87 Benzhydrazide,3-methoxy-N2-(4- C20H22N2O2 322 4.62 Phenylcyclohexylideno)-

11 25.23 Squalene C30H50 410 5.07

12 26.40 Ricinoleic acid C18H34O3 298 10.56

13 28.24 ç-Tocopherol C28H48O2 416 6.29

14 29.76 Vitamin-E C29H50O2 430 10.49

15 33.47 ά-Sitosterol C29H50O 414 19.38

Table 2: Components identified in the chloroform leaf extract of Pterospermum canescens Roxb.

Molecular Peak S. No. RT Name of the compound MW formula area %

1 8.71 Asarone C12H16O3 208 9.43

2 9.20 1H-Cycloprop[e] azulen-7-ol,decahydro- C15H24O 220 3.13 1,1,7-trimethyl-4-methylene-, [1ar-(1aά, 4 aά, 7 ά, 7 aά, 7 bά)]- (Syn: Sputhulenol)

Cont… Int. J. Chem. Sci.: 10(2), 2012 1033

Molecular Peak S. No. RT Name of the compound MW formula area %

3 14.21 3,7,7-Trimethyl-1-penta-1-3-dienyl-2- C14H20O 204 49.03 oxbicyclo[3.2.0]hept-3-ene

4 14.84 2-Allyl-4-methylphenol C10H12O 148 30.49

5 15.41 Phytol C20H40O 296 3.31

6 25.62 Squalene C30H50 410 4.60

Table 3: Components identified in the methanol leaf extract of Pterospermum canescens Roxb.

Molecular Peak S. No. RT Name of the compound MW formula area%

1 3.52 1-Butanol, 3-methyl- formate C6H12O2 116 0.99

2 4.32 1-Methyl-pyrrolidine-2-carboxylic acid C6H11NO2 129 1.02

3 5.99 Phthalic anhydride C8H4O3 148 0.17

4 9.45 Diethyl Phthalate C12H14O4 222 53.84

5 9.94 Ethyl á-d-glucopyranoside C8H16O6 208 0.69

6 10.98 4-[(1E)-3-Hydroxy-1-propenyl]-2- C10H12O3 180 2.33 methoxyphenol

7 11.93 Oxirane, tetradecyl- C16H32O 240 0.54

8 12.97 Hexadecanoic acid, methyl ester C17H34O2 270 0.32

9 13.52 n-Hexadecanoic acid C16H32O2 256 0.35

10 13.81 Hexadecanoic acid, ethyl ester C18H36O2 284 0.45

11 15.13 9,12-Octadecadienoic acid, methyl C19H34O2 294 0.13 ester, (E,E)-

12 15.22 9,12,15-Octadecadienoic acid, methyl C19H32O2 292 0.28 ester, (Z,Z,Z,)-

13 15.36 Phytol C20H40O 296 0.51

14 16.04 Linoleic acid ethyl ester C20H36O2 308 0.49 Cont… 1034 K. P. Jaiganesh and G. Arunachalam: GC-MS Analysis of….

Molecular Peak S. No. RT Name of the compound MW formula area%

15 16.13 Ethyl Oleate C20H38O2 310 0.54

16 16.49 Octadecanoic acid, ethyl ester C20H40 O2 312 0.34

17 17.95 Methyl ricinoleate C19H36O3 312 3.93

18 18.98 Ricinoleic acid C18H34O3 298 20.74

19 21.87 7-Methyl-z-tetradecen-1-ol acetate C17H32O2 268 2.10

20 26.55 Glyceryl monoricinoleate C21H40O5 372 10.25 Rel. intensity

m/z Fig. 4: Mass spectrum of Vitamin E Rel. intensity Rel.

m/z Fig. 5: Mass spectrum Linoleic acid ethyl ester Int. J. Chem. Sci.: 10(2), 2012 1035 Rel. intensity

m/z Fig. 6: Mass spectrum of 9-Octadecenoic acid, 12-hydroxy-, methyl ester, [R-(Z)]- Rel. intensity Rel.

m/z Fig. 7: Mass spectrum of asarone Rel. intensity Rel.

m/z Fig. 8: Mass spectrum of Phytol 1036 K. P. Jaiganesh and G. Arunachalam: GC-MS Analysis of…. Rel. intensity

m/z Fig. 9: Mass spectrum of ethyl oleate Rel. intensity Rel.

m/z Fig. 10: Mass spectrum of phthalic anhydride Rel. intensity Rel.

m/z Fig. 11: Mass spectrum of diethyl phthalate Int. J. Chem. Sci.: 10(2), 2012 1037 Rel. intensity

m/z Fig. 12: Mass spectrum of hexadecanoic acid, methyl ester Rel. intensity Rel.

m/z Fig. 13: Mass spectrum of n-hexadecanoic acid

ACKNOWLEDGEMENT

The authors acknowledge to the Director, CRD, PRIST University, Thanjavur, Tamil Nadu for constant support during this study. The authors are very much thankful to Dr. K. Alagusundaram, Director, Indian Institute of Crop Processing Technology (IICPT), Thanjavur, Tamil Nadu. Authors are thankful to the Chairman, Dr. K. Varadharaajen, B.A., B.L., and Vice Chairman, Mr. John Ashok, M.A., B.L., for providing all the facilities to carry out this work. 1038 K. P. Jaiganesh and G. Arunachalam: GC-MS Analysis of….

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Revised : 15.02.2012 Accepted : 16.02.2012